WO2011112106A2 - Urease test for detecting helicobacter pylori bacteria - Google Patents

Urease test for detecting helicobacter pylori bacteria Download PDF

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Publication number
WO2011112106A2
WO2011112106A2 PCT/PL2011/000029 PL2011000029W WO2011112106A2 WO 2011112106 A2 WO2011112106 A2 WO 2011112106A2 PL 2011000029 W PL2011000029 W PL 2011000029W WO 2011112106 A2 WO2011112106 A2 WO 2011112106A2
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WO
WIPO (PCT)
Prior art keywords
urea
well
helicobacter pylori
indicator
test
Prior art date
Application number
PCT/PL2011/000029
Other languages
French (fr)
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WO2011112106A3 (en
Inventor
Stanisław KAFEL
Original Assignee
Kafel Stanislaw
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Publication date
Application filed by Kafel Stanislaw filed Critical Kafel Stanislaw
Publication of WO2011112106A2 publication Critical patent/WO2011112106A2/en
Publication of WO2011112106A3 publication Critical patent/WO2011112106A3/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/58—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving urea or urease
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/48742—Determining urea by measuring the volume of a gas
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/046—Function or devices integrated in the closure
    • B01L2300/047—Additional chamber, reservoir
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/069—Absorbents; Gels to retain a fluid
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/205—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Campylobacter (G)

Definitions

  • the subject of the invention is urease test for detecting Helicobacter pylori bacteria. They may cause inflamatory states and ulcers in the stomach and duodenum.
  • the test has application in medical diagnostics.
  • the test for dectecting Helicobacter pylori invented by the same author is known, and it has been protected by Polish Patent No.192 707. That test consists of two elements separated from each other and not contacting each other. These elements are made of filter paper, one them being the urea element. The other one is the indicator element which is impregnated with the indicator solution, preferably phenol red and eventually also with a buffer and potassium sorbate. The specimen examined will have contact with the element impregnated with urea. This specimen which is taken from the patient's organism is being placed on the urea element and it is being slightly pressed to squeeze out the tissue juice.
  • Helicobacter pylori is present in the sample - urease enzyme produced by this microorganism decomopses the urea with which the filter paper is impregnated.
  • the originating here gaseous ammonium penetrates into the indicator element impregnated with the indicator, preferably phenol red and eventually also with a buffer and protassium sorbate.
  • the ammonium causes the pH change of the indicator towards alkaline direction which is manifested by change of the colour of the indicator element from yellow to amaranth.
  • the amaranth colour of the indicator element means that in tissue specimen taken from the patient's organism Helicobacter pylori presence has been confirmed.
  • the hitherto used test according to the procedure protected with the patent No. 192 707 was characteristic by that the urea element and the indicator element were separated from each other by impermeable plastic foil or the air layer to avoid the false positive results. That made difficult the penetration of the gaseous ammonium to the indicator element and delayed receiving result on the test. Prolonged time of the test reaction made the period of waiting for the test result longer which created inconvenience for the patient and for the medical personnel.
  • the aim of this invention is the elaboration of the test construction which will enable elimination of the present inconveniences by shortening the time necessary for obtaining the test result. It will improve the work of the health service.
  • the test for detecting Helicobacter pylori is in the form of transparent, preferably rectangular plate with overmeasures along the longer sides.
  • a well preferably circular, which from the front side looks like a convexity.
  • the reagent elements made of the filter paper.
  • the urea element is impregnated with the urea solution and the indicator element which is placed at the bottom is impregnated with the indicator solution, preferably phenol red.
  • Direct contact with the tissue collected from the organism which is to be examined has the urea element on which the tissue is being placed.
  • the essence of the invention is such that between the urea element and the indicator element an open-work element is placed which enables quick penetration of the gasepus ammonium from the urea element to the indicator element.
  • This open-work element has preferably the shape of a die made of thin plastic mesh or of other material enabling fast and free flow of ammonium from the urea element to the indicator element.
  • the gaseous ammonium reacting with the indicator with which the filter paper of the indicator element is impregnated causes change of its colour from yellow to amaranth.
  • the disc of the filter paper of the indicator element with the changed colour which may be seen from the bottom of the well and also from the front side of the transparent plate enables to state whether in the specimen collected from the organism Helicobacter pylori is present. Intensity of the infection with this bacterium enables making comparison of colour of the indicator element with colours of models located on the label near the well which is seen from the front of the plate as a convexity.
  • fig.1 - showes general sight of the test together with the bended back self-sticking label
  • fig.2 - illustrates transversal profile of the test A-A together with the detail
  • fig.3 - the disc of the urea element
  • fig. 4- the disc of the open -work element
  • fig.5 - the disc of the indicator element.
  • the urease test for detecting Helicobacter pylori bacteria is in the form of transparent, rectangular plate 1 with overmeasures 2 along the longer sides.
  • a well 3 with the circular outline which is visible from the front side as a convexity.
  • the foil 4 To the plate 1 partly from the front side and partly from the bottom attached is the foil 4.
  • the disc 5 To the internal side of the foil 4 the disc 5 with the outline corresponding to the outline of the well is attached.
  • the reagent elements made of the filter paper impregnated with the reagents.
  • the urea element 6 is impregnated with urea soution and the indicator element 7 with the phenol red solution.
  • the open-work element 8 enables quick penetration of the gaseous ammonium which appears as a result of urease enzyme reaction, which the enzyme is produced by Helicobacter pylori bacteria present in the tissue collected from the patient's organism placed on the urea element 6, pressed down with the disc 5 together with the foil 4 to squeeze the tissue juice.
  • the gaseous ammonium penetrating to the filter paper indicator element 7 which is at the bottom of the well 3 comes into reaction with the phenol red and induces appearance of amaranth colour of the indicator element 7.
  • the intensity of this colour is being compared with the models 9 which are present in front of the plate 1 on the label 4 creating possibility of estimation whether Helicobacter pylori is present and in what concentration in the collected sample.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Physics & Mathematics (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Hematology (AREA)
  • Food Science & Technology (AREA)
  • Urology & Nephrology (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Toxicology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The invention solves the problem of construction of the urease test for detecting Helicobacter pylori bacteria. The test is in the form of transparent plate (1) with overmeasures (2) and the well (3) which from the front is visible as a convexity. To the plate (1) is fixted selfsticking foil (4). In the well (3) there are the filter paper reagent elements. The urea element (6) is impregnated with the urea solution and the indicator element (T) with fenol red solution. Between the urea element (6) and the indicator element (T) there is the open-work element (8).

Description

Urease test for detecting Helicobacter pylori bacteria
The subject of the invention is urease test for detecting Helicobacter pylori bacteria. They may cause inflamatory states and ulcers in the stomach and duodenum. The test has application in medical diagnostics.
In the course of long term scientific investigations it was found that the above diseases may be caused by Helicobacter pylori bacteria. Characteristic feature of these bacteria as opposed to all other bacteria, is their ability of producing very big amount of urease - the enzym decomposing urea. In the course of urea decomposing, the ammonium and carbon dioxide is being produced and ammonium induces the pH raising in the environment where this reaction takes places. It was decided that this characteristic will be utilized in the urease tests for detecting the above mentioned Helicobacter pylori bacteria.
In all hitherto existing known urease tests the urea and the indicator and preferably phenol red are used and when urea is being decomposed by urease, phenol red turns its colour from yellow to amaranth. In the hitherto known urease tests the urea and the indicator used are being diluted in water or in agar gel, or they are in the filter paper impregnated with the solutions of these constituents.
The test for dectecting Helicobacter pylori invented by the same author is known, and it has been protected by Polish Patent No.192 707. That test consists of two elements separated from each other and not contacting each other. These elements are made of filter paper, one them being the urea element. The other one is the indicator element which is impregnated with the indicator solution, preferably phenol red and eventually also with a buffer and potassium sorbate. The specimen examined will have contact with the element impregnated with urea. This specimen which is taken from the patient's organism is being placed on the urea element and it is being slightly pressed to squeeze out the tissue juice. If Helicobacter pylori is present in the sample - urease enzyme produced by this microorganism decomopses the urea with which the filter paper is impregnated. The originating here gaseous ammonium penetrates into the indicator element impregnated with the indicator, preferably phenol red and eventually also with a buffer and protassium sorbate. The ammonium causes the pH change of the indicator towards alkaline direction which is manifested by change of the colour of the indicator element from yellow to amaranth. The amaranth colour of the indicator element means that in tissue specimen taken from the patient's organism Helicobacter pylori presence has been confirmed.
It is essential to mention that examination of the specimen callected from the patient's organism by the endoscopic method is being carried out on the test immediately after the specimen was obtained.
The hitherto used test according to the procedure protected with the patent No. 192 707 was characteristic by that the urea element and the indicator element were separated from each other by impermeable plastic foil or the air layer to avoid the false positive results. That made difficult the penetration of the gaseous ammonium to the indicator element and delayed receiving result on the test. Prolonged time of the test reaction made the period of waiting for the test result longer which created inconvenience for the patient and for the medical personnel. The aim of this invention is the elaboration of the test construction which will enable elimination of the present inconveniences by shortening the time necessary for obtaining the test result. It will improve the work of the health service.
According to the invention the test for detecting Helicobacter pylori is in the form of transparent, preferably rectangular plate with overmeasures along the longer sides. In the plate there is a well, preferably circular, which from the front side looks like a convexity. In the well there are placed the reagent elements made of the filter paper. The urea element is impregnated with the urea solution and the indicator element which is placed at the bottom is impregnated with the indicator solution, preferably phenol red. Direct contact with the tissue collected from the organism which is to be examined has the urea element on which the tissue is being placed.
The essence of the invention is such that between the urea element and the indicator element an open-work element is placed which enables quick penetration of the gasepus ammonium from the urea element to the indicator element. This open-work element has preferably the shape of a die made of thin plastic mesh or of other material enabling fast and free flow of ammonium from the urea element to the indicator element.
The gaseous ammonium reacting with the indicator with which the filter paper of the indicator element is impregnated causes change of its colour from yellow to amaranth. The disc of the filter paper of the indicator element with the changed colour which may be seen from the bottom of the well and also from the front side of the transparent plate enables to state whether in the specimen collected from the organism Helicobacter pylori is present. Intensity of the infection with this bacterium enables making comparison of colour of the indicator element with colours of models located on the label near the well which is seen from the front of the plate as a convexity.
The subject of the invention has been described on the enclosed illustration on which fig.1 - showes general sight of the test together with the bended back self-sticking label, fig.2 - illustrates transversal profile of the test A-A together with the detail, fig.3 - the disc of the urea element, fig. 4- the disc of the open -work element, and fig.5 - the disc of the indicator element.
The urease test for detecting Helicobacter pylori bacteria, according to the invention, is in the form of transparent, rectangular plate 1 with overmeasures 2 along the longer sides. In the plate 1 there is a well 3 with the circular outline which is visible from the front side as a convexity. To the plate 1 partly from the front side and partly from the bottom attached is the foil 4. To the internal side of the foil 4 the disc 5 with the outline corresponding to the outline of the well is attached. In the well 3 there are the reagent elements made of the filter paper impregnated with the reagents. The urea element 6 is impregnated with urea soution and the indicator element 7 with the phenol red solution. Between the urea element 6 and the indictor element 7 there is the open - work element 8 made of plastic mesh. The open-work element 8 enables quick penetration of the gaseous ammonium which appears as a result of urease enzyme reaction, which the enzyme is produced by Helicobacter pylori bacteria present in the tissue collected from the patient's organism placed on the urea element 6, pressed down with the disc 5 together with the foil 4 to squeeze the tissue juice. The gaseous ammonium penetrating to the filter paper indicator element 7 which is at the bottom of the well 3 comes into reaction with the phenol red and induces appearance of amaranth colour of the indicator element 7. The intensity of this colour is being compared with the models 9 which are present in front of the plate 1 on the label 4 creating possibility of estimation whether Helicobacter pylori is present and in what concentration in the collected sample.

Claims

Claims
1. The urease test for detecting Helicobacter pylori bacteria, which is in the form of transparetnt plate with overmeasures along the longer sides and with the well, preferably of the circular outline which is visible from the front side as a convexity, and to this plate from its front and from the bottom the selfstieking foil with a disc corresponding to the outline of the well is attached, in the well there are the filter paper reagent elements impregnated with the reagents where the urea element is impregnated with the urea solution and the indicator element with the phenol red solution, and eventually with a buffer and potassium sorbate, characterized in that between the urea element (6) and the indicator element (7) there is the open-work element (8).
2. The urease test according to claim 1, characterized in that the open-work element (8) has preferably the shape of a disc made of plastic mesh or of other material enabling fast and free flow of ammonium from the urea element (6) to the indicator element (7).
PCT/PL2011/000029 2010-03-10 2011-03-04 Urease test for detecting helicobacter pylori bacteria WO2011112106A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
PL390673A PL390673A1 (en) 2010-03-10 2010-03-10 Urease test for detection of Helicobacter pylori
PLP-390673 2010-03-10

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WO2011112106A2 true WO2011112106A2 (en) 2011-09-15
WO2011112106A3 WO2011112106A3 (en) 2011-12-08

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012051644A1 (en) * 2010-10-18 2012-04-26 Centre For Digestive Diseases Pty Limited Compostion and apparatus for detecting urease in gastric contents
CN102539424A (en) * 2011-12-23 2012-07-04 李雄 Rapid detection method of urease in tongue coating by using pH value-ethanol method and kit
DE202014101208U1 (en) 2013-03-26 2014-07-04 Strong Biotech Corp. Diagnostic device for Helicobacter pylori
CN105424931A (en) * 2015-04-04 2016-03-23 吉林双正医疗科技有限公司 Helicobacter pylori urease antibody IgM-IgG combination rapid test device and preparation method thereof
EP3009519A1 (en) * 2014-10-14 2016-04-20 AMA-Med Oy Test device, reactive element and arrangement for urease activity
CN110470663A (en) * 2019-08-21 2019-11-19 王冠男 For quickly detecting the reagent and kit of people's stomach Helicobacter pylori
RU2715405C1 (en) * 2018-12-24 2020-02-27 Общество с ограниченной ответственностью "Ассоциация Медицины и Аналитики" (ООО "АМА") Test system for determining bacterial fermentative activity by ammonia

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
PL192707B1 (en) 2001-03-20 2006-12-29 Kafel Stanislaw Novel solution for a test serving to detect urease produced by heliobacter pylori

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5877663A (en) * 1981-11-02 1983-05-11 Kyoto Daiichi Kagaku:Kk Method and implement for analysis of urea
CA1274757A (en) * 1985-05-17 1990-10-02 Barry James Marshall Compositions and methods for the detection of urease for the diagnosis of campylobacter pyloridis infection
US5314804A (en) * 1992-03-24 1994-05-24 Serim Research Corporation Test for Helicobacter pylori
US6998250B2 (en) * 2001-10-15 2006-02-14 Donald J. McMichael Method for detecting Helicobacter pylori
CN101748185A (en) * 2008-12-02 2010-06-23 实创国际生技股份有限公司 Detection test piece and detection method thereof for stomach Helicobacter pylori

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
PL192707B1 (en) 2001-03-20 2006-12-29 Kafel Stanislaw Novel solution for a test serving to detect urease produced by heliobacter pylori

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012051644A1 (en) * 2010-10-18 2012-04-26 Centre For Digestive Diseases Pty Limited Compostion and apparatus for detecting urease in gastric contents
CN102539424A (en) * 2011-12-23 2012-07-04 李雄 Rapid detection method of urease in tongue coating by using pH value-ethanol method and kit
DE202014101208U1 (en) 2013-03-26 2014-07-04 Strong Biotech Corp. Diagnostic device for Helicobacter pylori
EP3009519A1 (en) * 2014-10-14 2016-04-20 AMA-Med Oy Test device, reactive element and arrangement for urease activity
CN105424931A (en) * 2015-04-04 2016-03-23 吉林双正医疗科技有限公司 Helicobacter pylori urease antibody IgM-IgG combination rapid test device and preparation method thereof
CN105424931B (en) * 2015-04-04 2016-08-17 吉林双正医疗科技有限公司 Helicobacter Pylori urease antibody IgM, IgG associating device for fast detecting and preparation method thereof
RU2715405C1 (en) * 2018-12-24 2020-02-27 Общество с ограниченной ответственностью "Ассоциация Медицины и Аналитики" (ООО "АМА") Test system for determining bacterial fermentative activity by ammonia
WO2020139142A1 (en) 2018-12-24 2020-07-02 Общество с ограниченной ответственностью "Ассоциация Медицины и Аналитики" (ООО "АМА") Testing system for determining the enzymatic activity of bacteria on ammonia
EP3904497A4 (en) * 2018-12-24 2022-05-04 "Association of Medicine and Analytics" Co. Ltd (AMA Co. Ltd) Testing system for determining the enzymatic activity of bacteria on ammonia
CN110470663A (en) * 2019-08-21 2019-11-19 王冠男 For quickly detecting the reagent and kit of people's stomach Helicobacter pylori

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WO2011112106A3 (en) 2011-12-08

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